Microbial community analysis of a methane-oxidizing biofilm using ribosomal tag pyrosequencing

Kim Tae Gwan, Eun Hee Lee, Kyung Suk Cho

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25 Scopus citations


Current ecological knowledge of methanotrophic biofilms is incomplete, although they have been broadly studied in biotechnological processes. Four individual DNA samples were prepared from a methanotrophic biofilm, and a multiplex 16S rDNA pyrosequencing was performed. A complete library (before being de-multiplexed) contained 33,639 sequences (average length, 415 nt). Interestingly, methanotrophs were not dominant, only making up 23% of the community. Methylosinus, Methylomonas, and Methylosarcina were the dominant methanotrophs. Type II methanotrophs were more abundant than type I (56 vs. 44%), but less richer and diverse. Dominant non-methanotrophic genera included Hydrogenophaga, Flavobacterium, and Hyphomicrobium. The library was de-multiplexed into four libraries, with different sequencing efforts (3,915 - 20,133 sequences). Sørrenson abundance similarity results showed that the four libraries were almost identical (indices > 0.97), and phylogenetic comparisons using UniFrac test and P-test revealed the same results. It was demonstrated that the pyrosequencing was highly reproducible. These survey results can provide an insight into the management and/or manipulation of methanotrophic biofilms.

Original languageEnglish
Pages (from-to)360-370
Number of pages11
JournalJournal of Microbiology and Biotechnology
Issue number3
StatePublished - 3 Dec 2011


  • Biofilm
  • Community analysis
  • Methanotrophs
  • Pyrosequencing
  • Quantitative real-time PCR


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